Name: Time-Lapse Epifluorescentiemicroscopie Beeldvorming van Pseudomonas aeruginosa en Staphylococcus aureus Heterogene fenotypes (Video) | JoVE Journal
Uploaded: 2025-02-14T14:30:00.000+00:00
Duration: 7 min 44 s
Description: Leer een reproduceerbare time-lapse epifluorescentiemicroscopieprotocol voor het visualiseren van heterogene fenotypes in Pseudomonas aeruginosa en Staphylococcus aureus. Deze methode helpt onderzoekers overlevingsstrategieën van persistercellen te bestuderen.

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<meta charset="utf8"></head><body>Visualisatie van antibioticapersistentie bij bacteriële pathogenen

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<table><tbody><tr><td>BaSiC</td><td>GitHub</td><td>https://github.com/marrlab/BaSiC</td><td></td></tr><tr><td>Certified Molecular Biology Agarose</td><td>Biorad</td><td>1613101</td><td></td></tr><tr><td>Fiji-ImageJ</td><td>NIH</td><td>https://imagej.net/software/fiji/downloads</td><td></td></tr><tr><td>Interchangeable Coverglass Dish</td><td>Bioptechs</td><td>190310-35</td><td>35 mm ICD voor het bereiden van agarose pads; wordt geleverd met 30 mm (#1.5) dekvlakken</td></tr><tr><td>Lumencor Spectra 7 LED light engine</td><td>Lumencor</td><td>https://lumencor.com/products/spectra-light-engine</td><td>Spectra 7 LED light engine</td></tr><tr><td>MetaMorph</td><td>Molecular Devices</td><td>Premier versie 7.10.5</td><td></td></tr><tr><td>pco.edge 4.2 bi sCMOS camera</td><td>Excelitas</td><td>https://www.excelitas.com/product/pcoedge-42-bi-usb-scmos-camera</td><td>sCMOS camera</td></tr><tr><td>PeCon live cell incubatiekamer</td><td>peCon</td><td>https://www.pecon.biz/</td><td></td></tr><tr><td>Thomas Scientific Ronde dekglas, #1.5 dikte, 25 mm, 100 pack</td><td>Fisher Scientific</td><td>NC1272770</td><td>25 mm (#1.5) dekvlakken</td></tr><tr><td>Zeiss Axiovert 200M microscoop</td><td>Zeiss</td><td>https://www.zeiss.com/microscopy/en/products/light-microscopes/widefield-microscopes/axiovert-for-materials.html</td><td>Omgekeerd microscoop met Plan-Apochromat 63x/1.40 Oil Ph3 M27 objectief, Lumencor Spectra 7 LED light engine, en pco.edge 4.2 bi sCMOS camera (6.5 mm pixelgrootte)</td></tr></tbody></table>

time-lapse epifluorescentiemicroscopie beeldvorming van pseudomonas aeruginosa en staphylococcus aureus heterogene fenotypes

Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes

Time-Lapse Epifluorescentiemicroscopie Beeldvorming van Pseudomonas aeruginosa en Staphylococcus aureus Heterogene fenotypes

Antibiotic persistence is a phenomenon in which a small number of bacterial cells in a genetically susceptible population survive antibiotic treatment ...

time-lapse-epifluorescence-microscopy-imaging-pseudomonas-aeruginosa

Research

JoVE Journal

Biology

Biologie

Protocol beschrijft de toepassing van een flow cel-systeem voor de teelt en het analyseren van microbiële biofilms voor Confocale Laser Scanning Microscopie (CLSM).

Pseudomonas aeruginosa en Saccharomyces cerevisiae Biofilm in Flow cellen

Dit protocol biedt een stap-voor-stap procedure te controleren enkele cel gedrag van verschillende bacteriën in de tijd met behulp van geautomatiseerde fluorescentie time-lapse microscopie. Verder hebben we geven richtlijnen hoe de microscopie beelden te analyseren.

Live cell imaging van Bacillus subtilis En Streptococcus pneumoniae Met Automatisch Time-lapse Microscopie

Source: Chetrit, D., et al. Applying Live Cell Imaging and Cryo-Electron Tomography to Resolve Spatiotemporal Features of the Legionella pneumophila Dot/Icm Secretion System. J. Vis. Exp. (2020).This video demonstrates the preparation and imaging of fluorescently labeled pathogenic bacteria. Agarose pads are first prepared and positioned within an adhesive frame. A suspension of genetically modified bacteria expressing a fluorescent protein is applied to the pad to immobilize the cells. The...

Sample Preparation and Imaging of Fluorescently Labeled Bacteria

Source: Jackson, L., et al. Visualization of Pseudomonas aeruginosa within the Sputum of Cystic Fibrosis Patients. J. Vis. Exp. (2020)This video demonstrates the in situ visualization of Pseudomonas aeruginosa within chemically cleared cystic fibrosis sputum using fluorescent labeling and confocal microscopy to reveal bacterial localization and biofilm structure.

Visualization of Pseudomonas aeruginosa in Sputum Samples from Cystic Fibrosis Patients

This video demonstrates an in vitro technique to study the impact of antimicrobial treatment on Pseudomonas aeruginosa (Pa) aggregates. Fluorescently-labeled Pa cells are grown as aggregates, mimicking the growth pattern during infection. Upon challenging the aggregates with a sublethal concentration of the antibiotic colistin, the real-time assessment of the bacteria-antibiotic interaction is performed via fluorescence microscopy.

An In Vitro Technique to Study Antimicrobial Treatments on Bacterial Aggregates

We methode om het effect van PSMs en andere toxines uitgescheiden door...

Studeren Interacties van<em&gt; Staphylococcus aureus</em&gt; Met Neutrofielen door flowcytometrie en Time Lapse Microscopie

We beschrijven fixatie, paraffine insluiten en dunne vectorafbeeldingsbestanden technieken voor microbiële kolonie biofilms. In bereid monsters, kunnen biofilm substructuur en verslaggever expressiepatronen worden gevisualiseerd door microscopie.

Paraffine insluiten en dunne afdelen van microbiële kolonie Biofilms voor microscopische analyse

Dit live-bacteriële celbeeldvormingsprotocol maakt visualisatie mogelijk van interacties tussen meerdere bacteriële soorten op het eencellige niveau in de tijd. Time-lapse imaging maakt het mogelijk om elke bacteriesoort in monocultuur of cocultuur te observeren om interspecies interacties te ondervragen in bacteriële gemeenschappen met meerdere soorten, waaronder individuele celmotiliteit en levensvatbaarheid.

Kinetische visualisatie van single-cell interspecies bacteriële interacties

Synthetisch cystic fibrosis sputum medium (SCFM2) kan worden gebruikt in combinatie met zowel confocale laserscanmicroscopie als fluorescentie-geactiveerde celsortering om bacteriële aggregaten met hoge resolutie te observeren. Dit artikel beschrijft methoden om geaggregeerde populaties te beoordelen tijdens antimicrobiële behandeling als een platform voor toekomstige studies.

Tools voor de real-time beoordeling van een Pseudomonas aeruginosa-infectiemodel

Antibioticapersistentie beschrijft het vermogen van kleine subpopulaties binnen een gevoelige isogene populatie om tijdelijk hoge doses bacteriedodende antibiotica te verdragen. Het huidige protocol combineert benaderingen om het fenotype van antibioticapersistentie op moleculair en cellulair niveau te karakteriseren na blootstelling van Escherichia coli aan dodelijke doses ofloxacine.

Populatie- en eencellige analyse van antibioticapersistentie in Escherichia coli

Leer een reproduceerbare time-lapse epifluorescentiemicroscopieprotocol voor het visualiseren van heterogene fenotypes in Pseudomonas aeruginosa en Staphylococcus aureus. Deze methode helpt onderzoekers overlevingsstrategieën van persistercellen te bestuderen.

Time-Lapse Epifluorescentiemicroscopie Beeldvorming van Pseudomonas aeruginosa en Staphylococcus aureus Heterogene fenotypes

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